Osteoblastic differentiation of bone marrow mesenchymal stromal cells in Bruck Syndrome. Issue 1 (December 2016)

• Record Type: Journal Article
• Authors: Kaneto, Carla; Lima, Patrícia; Zanette, Dalila; Oliveira, Thiago; de Assis Pereira, Francisco; Lorenzi, Julio; dos Santos, Jane; Prata, Karen; Neto, João; de Paula, Francisco; Silva, Wilson
• Is Part Of: BMC medical genetics. Volume 17:Issue 1(2016)
• Page Start: 1
• Page End: 9
• Publication Date: 2016-12
• Subjects: Bruck syndrome -- Osteogenesis Imperfecta -- Bone marrow mesenchymal stromal cell -- Osteogenic differentiation -- Gene expression; Medical genetics -- Periodicals; 616.04205
• DOI: 10.1186/s12881-016-0301-7
• Languages: English
• ISSNs: 1471-2350

Abstract

Abstract Background Osteogenesis Imperfecta (OI) (OMIM %259450) is a heterogeneous group of inherited disorders characterized by increased bone fragility, with clinical severity ranging from mild to lethal. The majority of OI cases are caused by mutations inCOL1A1 orCOL1A2 . Bruck Syndrome (BS) is a further recessively-inherited OI-like phenotype in which bone fragility is associated with the unusual finding of pterygia and contractures of the large joints. Notably, several studies have failed to show any abnormalities in the biosynthesis of collagen 1 in BS patientes. Evidence was obtained for a specific defect of the procollagen telopeptide lysine hydroxylation in BS, whereas mutations in the genePLOD2 have been identified. Recently, several studies described FKBP10 mutations in OI-like and BS patients, suggesting thatFKBP10 is a bonafide BS locus. Methods We analyzed the coding region and intron/exon boundaries ofCOL1A1, COL1A2, PLOD2 andFKBP10 genes by sequence analysis using an ABI PRISM 3130 automated sequencer and Big Dye Terminator Sequencing protocol. Mononuclear cells obtained from the bone marrow of BS, OI patients and healthy donors were cultured and osteogenic differentiation was induced. The gene expression of osteoblast specific markers were also evaluated during the osteoblastic differentiation of mesenchymal stem cell (MSC) by qRT-PCR using an ABI7500 Sequence Detection System. Results No mutations inCOL1A1, COL1A2 orPLOD2 were found in BS patient. We found a homozygous 1-base-pair duplication (c.831dupC) that is predicted to produce a translational frameshift mutation and a premature protein truncation 17 aminoacids downstream (p.Gly278ArgfsX95). The gene expression of osteoblast specific markersBGLAP, COL1A1, MSX2, SPARC andVDR was evaluated by Real Time RT-PCR during differentiation into osteoblasts and results showed similar patterns of osteoblast markers expression in BS and healthy controls. On the other hand, when compared with OI patients, the expression pattern of these genes was found to be different. Conclusions Our work suggests that the gene expression profiles observed during mesenchymal stromal cell differentiation into osteoblast are distinct in BS patients as compared to OI patients. The present study shows for the first time that genes involved in osteogenesis are differentially expressed in BS and OI patients.